Integrated Systems for the Analysis of Circulating Multiple Myeloma Cells (CMMCs)

 描述(申请人提供)：多发性骨髓瘤(MM)是一种浆细胞恶性肿瘤，约占恶性肿瘤的1%，占所有血液系统肿瘤的15%。根据美国癌症协会的数据，2014年美国估计新增24050例确诊病例，死亡人数估计为11090人。未确定意义的单克隆性伽马病(MGUS)每年有1%的风险进展为明显的恶性肿瘤，最常见的是MM，在50岁以上的人中约有3%发现，患病率随着年龄的增加而增加。由于没有明确的诊断试验来监测MGUS的恶性进展，目前监测到的一系列因素包括血清游离轻链比率的变化、血清M蛋白水平的升高和骨髓浆细胞百分比的升高。最近的研究表明，通过免疫fl荧光显微镜或flow细胞仪从骨髓活检组织中获得的循环多发性骨髓瘤细胞(CMMC)对于MM的诊断、预后和治疗反应是有用的。然而，确保骨髓活检对患者来说是痛苦的，因为禁止频繁的检测是监测疾病进展、药物反应和/或复发所必需的。开发用于直接从外周血中选择和分析CMMC的新仪器和技术对于改善MM患者或其早期疾病阶段的管理至关重要。基于从第一阶段类型研究中获得的大量初步数据，该第二阶段项目专注于开发一种综合系统，该系统使用一次性流体盒和相关的仪器外围设备，直接从全血中快速和完全自动化地处理CMMC，从而消除了对骨髓活检的需要，从而允许更频繁的测试，特别是MGUS的测试。该仪器将能够以80%的回收率和80%的纯度选择CMMC，释放选定的细胞并对它们进行电子计数，并准备用于FISH分析或免疫表型分析。该系统还可以卸载选定的CMMC进行分子图谱分析，例如搜索KRAS突变。仪器的操作将是自动化的，总体样本到答案的处理时间约为1.5小时。该系统将接受BioFluidica工作人员的内部验证以及北卡罗来纳大学教堂山分校合作者的外部验证。虽然MM将被用作该仪器的临床性能验证的案例研究，但通过编程在正确选择的抗体中搜索相关的血液生物细胞，它可以很容易地用于不同的癌症相关疾病。

 DESCRIPTION (provided by applicant): Multiple myeloma (MM) is a plasma cell malignancy and comprises about 1% of malignant tumors and 15% of all hematopoietic neoplasms. According to the American Cancer Society, in 2014 an estimated 24,050 new cases were diagnosed in the U.S. with the estimated deaths being 11,090. Monoclonal gammopathy of undetermined significance (MGUS) is associated with a 1% per year risk of progression to overt malignancy, most commonly MM and is found in approximately 3% of individuals over 50 with the prevalence increasing with age. Without a clear diagnostic test to monitor MGUS malignancy progression, a range of factors are currently monitored including changes in serum free light chains ratio, elevated serum M protein level and high bone marrow plasma cell percentage . Recent studies have demonstrated that circulating multiple myeloma cells (CMMCs) secured from bone marrow biopsies measured by immuno-fluorescence microscopy or flow cytometry are useful for diagnosis, prognosis and measuring therapeutic response for MM. However, securing a bone marrow biopsy is painful to the patient prohibiting frequent testing required for monitoring disease progression, drug response and/or recurrence. Development of novel instrumentation and techniques for CMMC selection and analysis directly from peripheral blood is essential to improve the management of patients with MM or its early disease stages. Building on extensive preliminary data secured from a Phase I type study, this Phase II project is focused on the development of an integrated system that uses a disposable fluidic cartridge and the associated instrument peripherals for the rapid and fully automated processing of CMMCs directly from whole blood obviating the need for bone marrow biopsies and thus, allowing for more frequent testing especially those with MGUS. The instrument will be able to select CMMCs at >80% recovery and >80% purity, release the selected cells and electronically enumerate them and prepare them for FISH analysis or immunophenotyping. The system can also off-load the selected CMMCs for molecular profiling, such as searching for KRAS mutations. Operation of the instrument will be automated with an overall sample- to-answer processing time of ~1.5 h. The system will be subjected to both internal validation by BioFluidica staff as well as external validation by collaborators at the University of North Carolina at Chapel Hill. While MM will be used as a case study for clinical performance verification of the instrument, it can easily be utilized in different cancer-related diseases by programming in the correct selection antibody to search for the relevant blood-borne biological cells.